7 March 2014 Point spread function optimization for STORM using adaptive optics
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Proceedings Volume 8978, MEMS Adaptive Optics VIII; 89780I (2014); doi: 10.1117/12.2040390
Event: SPIE MOEMS-MEMS, 2014, San Francisco, California, United States
Abstract
Stochastic Optical Reconstruction Microscopy (STORM) requires a high Strehl ratio point spread function (PSF) to achieve high resolution, especially in the presence of background fluorescence. The PSF is degraded by aberrations caused by imperfections in the optics, the refractive index mismatch between the sample and coverslip, and the refractive index variations of the sample. These aberrations distort the shape of the PSF and increase the PSF width directly reducing the resolution of STORM. Here we discuss the use of Adaptive Optics (AO) to correct aberrations, maintaining a high Strehl ratio even in thick tissue. Because the intensity fluctuates strongly from frame to frame, image intensity is not a reliable measure of PSF quality, and the choice of a robust optimization metric is critical. We demonstrate the use of genetic algorithms with single molecule imaging for optimization of the wavefront and introduce a metric that is relatively insensitive to image intensity. We demonstrate the correction of the wavefront from measurements of single quantum dots.
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Kayvan Forouhesh Tehrani, Peter Kner, "Point spread function optimization for STORM using adaptive optics", Proc. SPIE 8978, MEMS Adaptive Optics VIII, 89780I (7 March 2014); doi: 10.1117/12.2040390; http://dx.doi.org/10.1117/12.2040390
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KEYWORDS
Wavefronts

Adaptive optics

Point spread functions

Genetic algorithms

Microscopy

Molecules

Signal to noise ratio

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